Agrisera/HY1 | Heme Oxygenase 1/AS12 2636/


productinformation
Background		HY1(HEMEOXYGENASE1) is aplastidhemeoxygenasenecessaryforphytochromechromophorebiosynthesisandforcouplingtheexpressionofsomenucleargenestothefunctionalstateofthechloroplast.Alternativenames:HEMEOXYGENASE1,HEMEOXYGENASE1,GENOMESUNCOUPLED2,HO1,ARABIDOPSISTHALIANAHEMEOXYGENASE1,HY6,HY1,ATHO1,HEMEOXYGENASE6,GUN2,REVERSALOFTHEDETPHENOTYPE4.
Immunogen		KLH-conjugatedsyntheticpeptidederivedfromArabidopsisthaliana HY1,UniProt: O48782,TAIR:At2g26670
Host		Rabbit
Clonality		Polyclonal
Clone
Purity		AffinitypurifiedseruminPBS,pH7.4.
Format		Lyophilized
Quantity		50µg
Reconstitution		Forreconstitutionadd50µl,ofsterilewater.
Storage		Storelyophilized/reconstitutedat-20°C;oncereconstitutedmakealiquotstoavoidrepeatedfreeze-thawcycles.Please,remembertospintubesbrieflypriortoopeningthemtoavoidanylossesthatmightoccurfromlyophilizedmaterialadheringtothecaporsidesofthetubes.
Testedapplications		WesternBlot(WB)
Relatedproducts		Collectionofantibodiesinvolvedinphotomorphogenesis Plantproteinextractionbuffer Secondaryantibodies
Additionalinformation

applicationinformation
Recommendeddilution		1:1000(WB)
Expected\|apparentMW		32kDa
Confirmedreactivity		Arabidopsisthaliana
Predictedreactivity		Brassicajuncea,Capsicumannuum,Corchoruscapsularis,Morusalba,Nelumbonucifera,Nicotianasylvestris,Nicotianatabacum,Noccaeacaerulescens,Populusbalsamifera,Populustremula,Solanumlycopersicum
Notreactivein
Additionalinformation
Selectedreferences		Tobeaddedwhenavailable,antibodyreleasedinJuly2017.

Applicationexample

Arabidopsisthalianawild-type(Col-0)andhy1-100mutantseedlingsweregrownon½MSagarplatessupplementedwith1%agar,withoutsucrose,for2ddarkand3dinWLc(100µmolm-2s-1)at22°C.100mgofcotyledontissuewascollectedfrom5doldseedlingsandextractedwith500µL2xLaemmlibufferwithoutbromophenolblue(0.125MTris-HCl,pH6.8;4%SDS;20%glyceroland5%2-mercaptoethanol),denaturedat99°Cfor5minusingthermomixer(Eppendorf),andcentrifugedfor5minat13,000rpmat4°C.Approximately100µgproteinfrom2mgofleafmaterialwasloadedperlane(ina10µLvolume)withappropriatereductionsfor0.5xand0.25x,respectively.Proteinswereseparatedona4%stackingand12%resolvingSDS-PAGEgelusingstandardTris/Glycinerunningbuffer(25mMTris,192mMglycine,0.1%SDS)andblottedfor1hat100Vtoa0.22µmnitrocellulosemembranefromLicor.Awettransferwasused,withtransferbuffercontaining:20%methanol,25mMTris,192mMglycine.BlotswerestainedwithPonceauSsolution(0.1%PonceauSin5%aceticacid)andwashedbrieflywithTBS,thenblockedwith5%(w/v)milkinTBSfor1hatroomtemperature(RT)withagitation.Blotswereincubatedintheprimaryantibodyatadilutionof1:2000overnightat4°Cwithagitationin5%milkinTBS-T.Theantibodysolutionwasremoved,theblotrinsedbriefly,andwashedsixtimesfor5mininTBS-TatRTwithagitation.Theblotwasincubatedinasecondaryantibodyprotectedfromlight(donkeyanti-rabbitIgGIRDye800CWfromLicor,925-32213)dilutedto1:20000in5%(w/v)milkinTBS-Tfor45minatRTwithagitation.Theblotwaswashedsixtimesfor5mininTBS-TatRTwithagitationandthreetimesfor2minwithTBStoremoveresidualTween.Forvisualization,theblotwasexcitedwiththe700nmand800nmchannels,withscanningintensity3and5,respectively,usingtheOdysseyinfra-redblotscannerfromLicor.

CourtesyofSylwiaKacprzakandDr.MatthewJ.Terry,UniversityofSouthampton,UnitedKingdom

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Agrisera/HY1 | Heme Oxygenase 1/AS12 2636/